重组结核杆菌热休克蛋白10对人成骨细胞增殖及骨代谢的调节

摘要

BACKGROUND:Mycobacterium tuberculosis heat shock protein 10 (r-Mt cpn10) is one of the main factors that cause bone tuberculosis dissolution and absorption as wel as inhibits the proliferation of osteoblasts. Receptor activator of nuclear factor kappa B ligand and osteoprotegerin are the important factors influencing bone metabolism. OBJECTIVE:To observe the effect of r-Mt cpn10 on human osteoblast proliferation, alkaline phosphatase secretion, expression of receptor activator of nuclear factor-kappa B ligand mRNA and osteoprotegerin mRNA. METHODS:Human bone marrow stromal cel s were induced to differentiate into osteoblasts, and osteoblasts at passage 3 were cultured with various concentrations of r-Mt cpn10 (0.1, 1, 10 mg/L). Osteoblasts cultured without r-Mt CPN10 were assigned as controls. RESULTS AND CONCLUSION:MTT assay results showed that, compared with control group, r-Mt cpn10 at different concentrations inhibited osteoblast proliferation and alkaline phosphatase secretion (P<0.05). RT-PCR analysis showed that, r-Mt cpn10 at different concentrations increased receptor activator of nuclear factor-kappa B ligand mRNA expression (P<0.01), and inhibited osteoprotegerin mRNA expression in a concentration-dependent manner (P<0.01). 10 mg/L r-Mt cpn10 exhibited the strongest effect (P<0.01). The r-Mt cpn10 can inhibit osteoblast proliferation and alkaline phosphatase activity, and it may influence bone metabolism by regulating the expression of receptor activator of nuclear factor-kappa B ligand mRNA and osteoprotegerin mRNA.%背景：结核杆菌热休克蛋白10是引起骨结核骨质溶解和吸收主要因素之一，抑制成骨细胞的增殖。核因子κB受体活化因子配体及骨保护素是影响骨代谢的重要指标。　　目的：观察重组结核杆菌热休克蛋白10对人成骨细胞增殖、碱性磷酸酶活性和核因子κB受体活化因子配体mRNA及骨保护素mRNA表达的影响。　　方法：将人骨髓基质干细胞定向诱导分化筛选为成骨细胞，取第3代细胞，用含质量浓度为0.1，1，10 mg/L的重组结核杆菌热休克蛋白10培养液培养，对照组成骨细胞正常培养。　　结果与结论：MTT 实验结果显示，与对照组相比，不同质量浓度重组结核杆菌热休克蛋白10抑制成骨细胞增殖和碱性磷酸酶活性(P<0.05)。RT-PCR实验显示，与对照组相比，不同质量浓度的重组结核杆菌热休克蛋白10均增加成骨细胞中核因子κB受体活化因子配体mRNA表达(P<0.01)，降低护骨素mRNA表达(P<0.01)，均呈剂量依赖性，质量浓度10 mg/L的重组结核杆菌热休克蛋白10作用最明显(P<0.01)。结果证实，重组结核杆菌热休克蛋白10抑制成骨细胞增殖和碱性磷酸酶活性，通过调节核因子κB 受体活化因子配体mRNA及骨保护素mRNA表达而影响骨代谢。